Comparison of gene expression profiles of miR-142-/- primary megakaryocytes and WT primary megakaryocytes

Whole fetal livers were collected from mouse fetuses at embryonic day 14.5 (E14.5), and single-cell suspensions were prepared by successive passage through 18-, 21 and 23-gauge needles. Fetal liver cells were maintained in Dulbecco modified Eagle medium (DMEM; Invitrogen) supplemented with 10% fetal bovine serum (FBS; Invitrogen), 100 U/ml penicillin, 100g/ml streptomycin, and 50ng/ml recombinant human thrombopoietin (TPO; Peprotech). After 5 days of culture, megakaryocytes were purified using a discontinuous bovine serum albumin gradient (BSA, SigmaAldrich; 3%, 1.5%, and 0%). Total RNA was isolated with TriReagent (MRC) following manufacturers instructions, and its quality was assessed with ND1000 Nanodrop (Peqlab) and on a 1.5% agarose gel.

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Chapnik E, Rivkin N, Mildner A, Beck G, Pasvolsky R, Metzl-Raz E, Birger Y, Amir G, Tirosh I, Porat Z, Israel LL, Lellouche E, Michaeli S, Lellouche JP, Izraeli S, Jung S, Hornstein E